rSJYB1 inhibits collagen type I protein expression in hepatic stellate cells via down-regulating activity of collagen α1 (I) promoter
rSJYB1 inhibits collagen type I protein expression in hepatic stellate cells via down-regulating activity of collagen α1 (I) promoter
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rSJYB1 通过下调胶原蛋白 α 1 (I) 启动子的活性抑制肝星状细胞中 I 型胶原蛋白的表达
DOI：
10.1111/jcmm.14271
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发表时间：
2019-05-01
影响因子：
5.3
通讯作者：
Duan, Yinong
中科院分区：
文献类型：
--
作者：
Chen, Liuting;Ji, Zhaodong;Duan, Yinong
YB1是肝纤维化的负性调节因子。我们想知道SJYB1是日本血吸虫YB1的同源蛋白，在体外是否对肝纤维化有作用。在细菌系统中表达重组SJYB1(RSJYB1)蛋白，并用Ni-NTA His-Bind树脂纯化。培养人肝星状细胞系LX-2，并用rSJYB1处理。用Western印迹和荧光素酶分析检测rSJYB1对LX-2细胞的作用。我们成功地在细菌系统中表达和纯化了SJYB1，纯化后的rSJYB1能被S感染的兔血清识别。蛋白质印迹分析显示，rSJYB1抑制I型胶原的表达，但对α-平滑肌肌动蛋白(α-SMA)的表达影响不大。进一步分析表明，rSJYB1抑制了胶原α1(I)(COL1A1)启动子的活性，并作用于COL1A1启动子的-1592/-1176区域。我们的研究结果表明，rSJYB1介导的抗纤维化作用包括抑制COL1A1启动子的活性，进而抑制肝星状细胞中I型胶原的表达。
YB1 is a negative regulator in liver fibrosis. We wondered whether SJYB1, a homologous protein of YB1 from Schistosoma japonicum, has an effect on liver fibrosis in vitro. Recombinant SJYB1 (rSJYB1) protein was expressed in a bacterial system and purified by Ni-NTA His-Bind Resin. A human hepatic stellate cell line, the LX-2 cell line, was cultured and treated with rSJYB1. The role of rSJYB1 on LX-2 cells was then analysed by Western blot and luciferase assay. We succeeded in expressing and purifying SJYB1 in a bacterial system and the purified rSJYB1 could be recognized by S japonicum-infected rabbit sera. Western bolt analysis showed that rSJYB1 inhibited the expression of collagen type I, but had little effect on alpha-smooth muscle actin (alpha-SMA). Further analysis revealed that rSJYB1 inhibited the activity of collagen alpha 1 (I) (COL1A1) promoter and functioned at -1592/-1176 region of COL1A1 promoter. Our data demonstrate that rSJYB1-mediated anti-fibrotic activity involves inhibiting the activity of COL1A1 promoter and subsequently suppressing the expression of collagen type I in hepatic stellate cells.